We are requesting funds to purchase a PERKIN-ELMER 7700 SEQUENCE DETECTOR/ABI PRISM 6700 AUTOMATED WORKSTATION to serve 10 independent Principal Investigators, their respective postdoctoral fellows, graduate students and research technicians (a group numbering nearly 30 individuals) at Georgetown University Medical Center. The recent introduction of real time, fluorescence based quantitative PCR (Q-PCR) as a rapid and sensitive technique for precise quantitation of nucleic acid templates now allows for detailed monitoring of gene expression. Until now, there was no easy path to reproducible Q-PCR results. Existing methods required serial dilutions and the analysis of multiple replicates, which involve numerous manual steps that introduced variability. The ABI PRISM 7700 Sequence Detector/6700 AUTOMATED WORKSTATION, along with a unique closed-tube assay, eliminate this variability. The 7700 system monitors PCR at every cycle. This real-time detection generates quantitative data based on the PCR at early cycles when PCR fidelity is the highest. In addition, the 7700 system has a linear dynamic range of at least five orders of magnitude, reducing the need for serial dilutions. Results are available just one minute after thermal cycling is complete. During the past year, Dr. Suarez-Quian did a sabbatical with Dr. C. Noguchi, Section Chief, Laboratory Chemical Biology, NIDDK, to learn how to apply real-time, Q-RT-PCR to reproductive tissues using the ABI 7700 system. He now wishes to purchase this instrument and introduce this technology to investigators at Georgetown to more efficiently fulfill and complete their specific aims of NIH sponsored RO1 grant applications. All ten members of this application wish to quantify specific gene transcripts from their respective tissues of interests. The ABI 7700 system will form the nucleus of the Molecular Biology Core of the Department of Cell Biology and help to foster a more efficient and precise means to complete research aims of the respective investigators.